Cotton is the most important crop for the production of fiber that plays a key role in economic and social affairs. The aim of the study was to evaluate the impact of biofield energy treatment on cotton seeds regarding its growth, germination of seedling, glutathione (GSH) concentration, indole acetic acid (IAA) content and DNA fingerprinting using simple sequence repeat (SSR) markers for polymorphism analysis. The seeds of cotton cv. Stoneville-2 (Gossypium hirsutum L.) was obtained from DNA Land Marks Inc., Canada and divided into two groups. One group was remained as untreated, while the other was subjected to Mr. Trivedi biofield energy and referred as treated sample. The growth-germination of cotton seedling data showed higher germination (82%) in biofield treated seeds as compared to the control (68%). The alterations in length of shoot and root of cotton seedling was reported in the treated sample with respect to untreated seeds. However, the endogenous level of GSH in the leaves of treated cotton was increased by 27.68% as compared to the untreated sample, which may suggest an improved immunity of cotton plant. Further, the plant growth regulatory constituent i.e. IAA concentration was increased by 7.39%, as compared with the control. Besides, the DNA fingerprinting data, showed polymorphism (4%) between treated and untreated samples of cotton. The overall results suggest that the biofield energy treatment on cotton seeds, results in improved overall growth of plant, increase germination rate, GSH and IAA concentration were increased. The study assumed that biofield energy treatment on cotton seeds would be more useful for the production of cotton fiber.